AZ3146 (AZ-3146; AZ3146) is a novel, potent and selective Mps1 kinase (Monopolar spindle1) inhibitor with potential anticancer activity. It inhibits Mps1 with an IC50 of ~35 nM, and is less potent against closely related kinases such as FAK, JNK1, JNK2, and Kit.

Physicochemical Properties

Molecular Formula	C24H32N6O3
Molecular Weight	452.55
Exact Mass	452.253
CAS #	1124329-14-1
Related CAS #	1124329-14-1
PubChem CID	56973724
Appearance	Off-white to light yellow solid powder
Density	1.3±0.1 g/cm3
Boiling Point	621.0±65.0 °C at 760 mmHg
Flash Point	329.4±34.3 °C
Vapour Pressure	0.0±1.8 mmHg at 25°C
Index of Refraction	1.631
LogP	2.76
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	33
Complexity	669
Defined Atom Stereocenter Count	0
InChi Key	YUKWVHPTFRQHMF-UHFFFAOYSA-N
InChi Code	InChI=1S/C24H32N6O3/c1-28-12-10-17(11-13-28)33-18-8-9-19(21(14-18)32-3)26-23-25-15-20-22(27-23)30(24(31)29(20)2)16-6-4-5-7-16/h8-9,14-17H,4-7,10-13H2,1-3H3,(H,25,26,27)
Chemical Name	9-cyclopentyl-2-(2-methoxy-4-(1-methylpiperidin-4-yloxy)phenylamino)-7-methyl-7H-purin-8(9H)-one
Synonyms	AZ3146; AZ 3146; AZ-3146
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro	AZ3146 inhibits human Mps1Cat in an in vitro kinase assay, with an IC50 of about 35 nM. Additionally, full-length Mps1 immunoprecipitated from human cells is potently inhibited from autophosphorylating by AZ3146 [1]. HCC cell growth can be inhibited by AZ3146, a TTK-specific kinase inhibitor. To test for cytotoxicity in vitro, SMMC-7721 and BEL-7404 cells were used. The results indicated that the IC50 values were 7.13 μM (BEL-7404) and 28.62 μM (SMMC-7721). For four days, both cells received additional treatment at IC50 concentrations. There was a noticeable reduction in cell proliferation [2]. HCT116 cells were grown for ten days at 0.8 μM (GI50) AZ3146 and for three weeks at 2 μM AZ3146. In cell viability assays, sixteen clones, designated AzR1-16, were isolated and cell lines were created. AzR3 and 4 had GI50 values of roughly 3 μM (4-fold resistance), while the remaining clones had GI50 values of roughly 9 μM (11-fold resistance). All of the clones were resistant to AZ3146-induced cell death. The parental cell line rapidly exited mitosis within 10 minutes when 2 μM AZ3146 was added, according to time-lapse microscopy analysis of mitosis [3].
ln Vivo	Regarding the effect of AZ3146 administration in vivo, the evidence should be provided by performing the study in human or mice or other animal models.
Animal Protocol
References	[1]. Sustained Mps1 activity is required in mitosis to recruit O-Mad2 to the Mad1-C-Mad2 core complex. J Cell Biol. 2010 Jul 12;190(1):25-34. [2]. TTK activates Akt and promotes proliferation and migration of hepatocellular carcinoma cells. Oncotarget. 2015 Oct 27;6(33):34309-20. [3]. Naturally Occurring Mutations in the MPS1 Gene Predispose Cells to Kinase Inhibitor Drug Resistance. Cancer Res. 2015 Aug 15;75(16):3340-54.

Solubility Data

Solubility (In Vitro)

DMSO: 28 mg/mL (61.9 mM) Water:<1 mg/mL Ethanol:91 mg/mL (201.1 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.52 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (5.52 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2097 mL	11.0485 mL	22.0970 mL
	5 mM	0.4419 mL	2.2097 mL	4.4194 mL
	10 mM	0.2210 mL	1.1049 mL	2.2097 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.